ABSTRACT
<p><b>OBJECTIVE</b>To observe the ultrastructural changes of HeLa cells in response to tubeimoside I (TBMS1) treatment and the protective effect of cyclosporine A (CsA), and explore the role of intrinsic apoptosis pathway in TBMS1-induced HeLa cell apoptosis.</p><p><b>METHODS</b>HeLa cells were treated with TBMS1 (10-50 micromo/L) alone or in combination with 2 micromol/L CsA for 12 and 24 h and observed with transmission electron microscope (TEM) for the ultrastructural changes of the cells.</p><p><b>RESULTS</b>TBMS1 induced apoptosis of HeLa cells in a concentration- and time-dependant manner. Under TEM, the treated cells progressively shrunk and the intercellular space widened with loss of microvillus, mitochondrial swelling, rough endoplasmic reticulum enlargement, chromatin condensation, nuclear shrinkage and nuclear pyknosis as TBMS1 concentration increased. At low concentrations, CsA offered partial protection of the mitochondria from TBMS1-induced damage whereas high-concentration CsA did not.</p><p><b>CONCLUSION</b>TBMS1 induces ultrastructural changes typical for apoptosis of the HeLa cells, which provides morphological evidence for the role of intrinsic apoptosis pathway in TBMS1-induced apoptosis.</p>
Subject(s)
Female , Humans , Apoptosis , Cell Nucleus , Cyclosporine , Pharmacology , Dose-Response Relationship, Drug , Endoplasmic Reticulum, Rough , HeLa Cells , Immunosuppressive Agents , Pharmacology , Microscopy, Electron, Transmission , Mitochondrial Swelling , Saponins , Pharmacology , Time Factors , Triterpenes , Pharmacology , Uterine Cervical Neoplasms , PathologyABSTRACT
<p><b>OBJECTIVE</b>To investigate a possibility of repairing damaged brain by intracerebroventricular transplantation of neural stem cells (NSCs) in the adult mice subjected to glutamate-induced excitotoxic injury.</p><p><b>METHODS</b>Mouse NSCs were isolated from the brains of embryos at 15-day postcoitum (dpc). The expression of nestin, a special antigen for NSC, was detected by immunocytochemistry. Immunofluorescence staining was carried out to observe the survival and location of transplanted NSCs. The animals in the MSG + NSCs group received intracerebroventricular transplantation of NSCs (approximately 1.0 x 10(5) cells) separately on day 1 and day 10 after 10-d MSG exposure (4.0 g/kg per day). The mice in control and MSG groups received intracerebroventricular injection of Dulbecco's minimum essential medium (DMEM) instead of NSCs. On day 11 after the last NSC transplantation, the test of Y-maze discrimination learning was performed, and then the histopathology of the animal brains was studied to analyze the MSG-induced functional and morphological changes of brain and the effects of intracerebroventricular transplantation of NSCs on the brain repair.</p><p><b>RESULTS</b>The isolated cells were Nestin-positive. The grafted NSCs in the host brain were region-specifically survived at 10-d post-transplantation. Intracerebroventricular transplantation of NSCs obviously facilitated the brain recovery from glutamate-induced behavioral disturbances and histopathological impairs in adult mice.</p><p><b>CONCLUSION</b>Intracerebroventricular transplantation of NSCs may be feasible in repairing diseased or damaged brain tissue.</p>
Subject(s)
Animals , Mice , Cell Count , Disease Models, Animal , Embryo, Mammalian , Glutamic Acid , Toxicity , Injections, Intraventricular , Methods , Intermediate Filament Proteins , Metabolism , Mice, Inbred Strains , Nerve Tissue Proteins , Metabolism , Nestin , Neurons , Physiology , Neurotoxicity Syndromes , Pathology , General Surgery , Stem Cell Transplantation , Methods , Stem Cells , Physiology , Time FactorsABSTRACT
<p><b>OBJECTIVE</b>To investigate the role of mitochondria in TBMS1-medited apoptosis of human cervical carcinoma HeLa cell line.</p><p><b>METHOD</b>Mitochondrial transmembrane potentia (deltapsim) was assayed by flow cytometry. Apoptotic induction by TBMS1 was determined by gel electrophoresis of fragmented DNA. Cytochrome c (Cyt c) was detected by Western blotting.</p><p><b>RESULT</b>The results showed that TBMS1 induced apoptosis in HeLa cells, that TBMS1 decreased deltapsim and facilitated Cyt c release, and that TBMS1 induced apoptosis of HeLa cells dose-and time-dependently in accordance with increase of cytosolic Cyt c. TBMS1 had direct effect on isolated rat mitochondria which induced a time- and dose-dependent release of Cyt c from mitochondria. The results also showed that CsA protected partly HeLa cells from the effect of TBMS1.</p><p><b>CONCLUSION</b>The mitochondrial apoptosis pathway has effects on TBMS1 induced human cervical carcinoma HeLa cell line apoptosis.</p>